Traditional Chinese medicine plaster for treating fusobacterium difficile and reconstitution of flora and preparation method of traditional Chinese medicine plaster

Abstract

The invention discloses a traditional Chinese medicine plaster for adjuvant therapy of clostridium difficile infection and reconstruction of intestinal flora balance and a preparation method thereof, and relates to the technical field of traditional Chinese medicine plasters, the traditional Chinese medicine plaster comprises a base material layer, an ointment and a protective layer, the ointment is prepared from effective components, a penetrant, an excipient, an adsorbent and deionized water; the effective components are prepared from rhizoma atractylodis macrocephalae volatile oil, cortex magnoliae officinalis volatile oil, folium artemisiae argyi volatile oil, a rhizoma coptidis extract, a herba taraxaci extract, a radix paeoniae alba extract and a poria cocos extract; according to the present invention, the accurate intervention of the intestinal microenvironment is achieved through the umbilical region targeting drug delivery and the combination of the dual mechanisms of the transdermal absorption and the acupoint stimulation, the drug action time is prolonged through the adsorbent porous slow-release structure, the flora balance is synchronously adjusted, the pathogenic bacterium proliferation is inhibited, and the limitation of the traditional oral administration or injection therapy is broken through; an efficient and convenient traditional Chinese medicine external treatment solution is provided for the drug-resistant clostridium difficile infection.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine plasters, and particularly relates to a traditional Chinese medicine plaster for assisting in the treatment of Clostridioides difficile infection and reconstructing the balance of intestinal flora, and a preparation method thereof. Background Art

[0002] Clostridioides difficile (formerly known as Clostridium difficile) infection (CDI) is a serious intestinal disease mainly caused by toxins produced by Clostridioides difficile, which can lead to a series of clinical symptoms ranging from mild diarrhea to pseudomembranous colitis, toxic megacolon, and even death. In recent years, with the widespread use of antibiotics, the incidence and recurrence rate of Clostridioides difficile infection have shown an upward trend, and it has become an important nosocomial infection and public health problem globally.

[0003] Clostridioides difficile infection mainly occurs in the hospital environment, and the vast majority is exogenous infection. The hospital environment is the main source of Clostridioides difficile. This bacterium can form spores and survive for a long time in adverse environments, increasing the difficulty of infection control. In particular, the elderly, immunocompromised individuals, those who have long-term use of antibiotics, patients with renal failure, inflammatory bowel disease, or chronic liver disease are all high-risk groups.

[0004] Currently, the main treatment methods for Clostridioides difficile infection include:

[0005] Antibiotic treatment: Vancomycin, metronidazole, etc. are first-line drugs for the treatment of CDI. However, antibiotic treatment has obvious limitations, including high treatment failure rate, high recurrence rate (about 20% - 30% of patients will experience recurrence), and the problem of potentially further disrupting the balance of intestinal flora.

[0006] Fecal microbiota transplantation: For patients with recurrent CDI, fecal microbiota transplantation is an effective treatment method, but there are problems such as complex operation, low acceptance, and potential risks.

[0007] Probiotics: As an adjuvant treatment method, but the efficacy is still controversial.

[0008] Comprehensive Western medicine treatment: It includes stopping the use of antibiotics that may cause CDI, replenishing electrolytes, and using probiotics.

[0009] The main problems faced by existing treatment methods are high recurrence rate and treatment failure rate, and long-term use of antibiotics may lead to an increase in drug resistance, further exacerbating the treatment difficulty. In addition, existing treatments are mostly systemic medications, which may have unnecessary effects on tissues and organs outside the intestine and are difficult to maintain an effective local drug concentration in the intestine.

[0010] Traditional Chinese medicine has a long history and rich experience in the treatment of digestive system diseases. According to traditional Chinese medicine theory, Clostridium difficile infection belongs to the categories of "diarrhea" and "dysentery", and is related to factors such as internal accumulation of damp-heat and weakness of the spleen and stomach.

[0011] However, at present, traditional Chinese medicine is mainly administered orally in the treatment of CDI, which has problems such as low bioavailability and insufficient local drug concentration in the intestine. At the same time, there is a lack of traditional Chinese medicine topical preparations for the treatment of Clostridium difficile infection, especially preparations that can directly act on the colon.

[0012] As a traditional administration method, traditional Chinese medicine plasters have the advantages of local medication, sustained release effect, and convenient use. However, existing traditional Chinese medicine plasters are mostly used for diseases such as trauma and rheumatic pain, and there is less research on plaster preparations for visceral organ diseases, especially intestinal infectious diseases.

[0013] Developing a traditional Chinese medicine plaster for the treatment of Clostridium difficile infection can directly act on the intestinal lesion site through transdermal absorption, avoid the first-pass effect of oral administration, improve the bioavailability of the drug, and reduce systemic adverse reactions at the same time, which has important clinical application value. Summary of the Invention

[0014] In order to solve the deficiencies of the existing technology, the purpose of the present invention is to provide a traditional Chinese medicine plaster for adjuvant treatment of Clostridium difficile infection and reconstruction of intestinal flora balance and its preparation method. The present invention targets drug delivery through the umbilicus (Shenque acupoint), combines the dual mechanisms of transdermal absorption and acupoint stimulation to achieve precise intervention in the intestinal microenvironment, extends the drug action time through the porous slow-release structure of the adsorbent, synchronously regulates the flora balance and inhibits the proliferation of pathogenic bacteria, breaks through the limitations of traditional oral or injection therapies, and provides an efficient and convenient external treatment solution of traditional Chinese medicine for drug-resistant Clostridium difficile infection.

[0015] In order to achieve the above purpose, the present invention adopts the following technical scheme:

[0016] A traditional Chinese medicine plaster for adjuvant treatment of Clostridium difficile infection and reconstruction of intestinal flora balance, comprising a substrate layer, a medicinal paste, and a protective layer. The medicinal paste is made of 1-6 wt% active ingredient, 0.5-2 wt% penetrant, 0.5-2 wt% excipient, 40-52 wt% adsorbent, and 38-58 wt% deionized water; the active ingredient is made of the following components in parts by weight: 4-5 parts of Atractylodes macrocephala volatile oil, 3-6 parts of Magnolia officinalis volatile oil, 1-3 parts of Artemisia argyi volatile oil, 16-24 parts of Coptis chinensis extract, 8-12 parts of Taraxacum mongolicum extract, 12-18 parts of Paeonia lactiflora extract, and 12-20 parts of Poria cocos extract.

[0017] Preferably, the preparation method of the traditional Chinese medicine volatile oil is as follows: Take Atractylodes macrocephala, Magnolia officinalis, and Artemisia argyi, crush them, add them to deionized water for soaking, reflux extraction, collect the volatile oil, respectively obtain the volatile oil of Atractylodes macrocephala, the volatile oil of Magnolia officinalis, and the volatile oil of Artemisia argyi, and obtain the residue of the volatile substances for standby.

[0018] After moderately crushing the medicinal materials such as Atractylodes macrocephala, microcracks are generated in their cell walls due to mechanical crushing. When soaked in water, the medicinal materials absorb water and swell, further softening the cell structure; during the reflux process, water molecules penetrate into the cells to form an azeotropic system with the volatile oil. Under continuous heating, the volatile oil components are co-distilled with water vapor due to the reduction of vapor pressure, and after condensation, the layer separation is achieved by using the immiscibility of oil and water. The double cell wall breaking (mechanical crushing + hydration swelling) of the medicinal materials in this process promotes the release of volatile oil, while the physical mass transfer characteristics of steam distillation selectively extract thermosensitive volatile components, and the chemical structures of active substances such as terpenoids and phenylpropanoids are completely retained.

[0019] Preferably, the traditional Chinese medicine is crushed into 40 - 80 meshes, the weight of deionized water is 10 - 20 times that of the traditional Chinese medicine, soaked for 30 - 60 min, and the reflux extraction temperature is 100 °C.

[0020] Preferably, the preparation method of the traditional Chinese medicine extract is as follows: Take Coptis chinensis, Taraxacum mongolicum, Paeonia lactiflora, and Poria cocos, add them to ethanol, heat under reflux, combine the extracts, filter, recover ethanol under reduced pressure from the filtrate, and concentrate it into a thick paste, respectively obtain the extract of Coptis chinensis, the extract of Taraxacum mongolicum, the extract of Paeonia lactiflora, and the extract of Poria cocos, and obtain the residue of the extracts for standby.

[0021] When the medicinal materials such as Coptis chinensis are extracted with ethanol solution, medium-polar ethanol molecules selectively dissolve active components such as flavonoids and alkaloids based on the principle of similar solubility; during the heating process, ethanol penetrates the cell membrane to generate an osmotic pressure difference, promoting the effective components in the cells to diffuse to the solvent phase through the concentration gradient, and at the same time, the thermal motion enhances the molecular diffusion rate; the fractional reflux extraction breaks the solid-liquid two-phase distribution equilibrium successively through the principle of dynamic equilibrium, improving the solute migration efficiency; in the reduced pressure concentration stage, the low boiling point characteristic of ethanol is used for preferential gasification separation, avoiding the destruction of thermosensitive components at high temperature, and finally forming a thick paste system rich in polar-medium polar active molecules, and the functional components such as antibacterial and anti-inflammatory are completely retained.

[0022] Preferably, the ethanol concentration is 60 - 80 wt%, the weight of ethanol is 8 - 15 times that of the traditional Chinese medicine; heat under reflux and extract 2 times, each time for 1 - 2 h.

[0023] Atractylodes macrocephala: Its nature and flavor are bitter, sweet, and warm, and it belongs to the spleen and stomach meridians. It has the effects of invigorating qi and strengthening the spleen, drying dampness and promoting diuresis, and securing the exterior and stopping sweating, and is mainly used for treating abdominal distension and diarrhea caused by spleen deficiency and dampness. Its volatile oil contains sesquiterpenoid components such as atractylone and atractylenolide, which can destroy the sterol structure of the cell membrane of Clostridium difficile and inhibit the proliferation of the bacteria.

[0024] Magnolia officinalis: bitter, pungent and warm, it enters the spleen, stomach, lung and large intestine meridians, has the functions of drying dampness and eliminating phlegm, lowering qi and eliminating fullness, and is targeted at dampness blocking the middle jiao, causing abdominal distension, vomiting and diarrhea. The biphenyl lignans such as magnolol and honokiol in the volatile oil block the release of toxins by inhibiting the activity of bacterial β-glucosidase.

[0025] Mugwort: bitter, pungent and warm, it enters the liver, spleen and kidney meridians, can warm the meridians to stop bleeding, dispel cold and relieve pain, and is mainly used to treat cold-induced bleeding and skin eczema. The volatile oil contains monoterpenoids such as eucalyptol and camphor, which can interfere with the formation of bacterial biofilms and promote transdermal absorption of drugs.

[0026] Coptis chinensis: bitter and cold, enters the spleen, stomach, liver and gallbladder meridians, clears away heat and dampness, purges fire and detoxifies, and is specially used to treat damp-heat, fullness, and vomiting. Isoquinoline alkaloids such as berberine in the extract can inhibit bacterial nucleic acid metabolism and downregulate the expression of inflammatory factors.

[0027] Dandelion: bitter, sweet and cold, it enters the liver and stomach meridians, clears away heat and detoxifies, reduces swelling and resolves nodules, and is good at treating heat-toxic carbuncle. The extract contains flavonoids such as luteolin, which can reduce inflammatory infiltration of the intestinal mucosa.

[0028] White peony root: bitter, sour and slightly cold, enters the liver and spleen meridians, nourishes blood and yin, softens the liver and relieves pain, used for abdominal pain caused by blood deficiency. Monoterpene glycosides such as paeoniflorin extract can regulate immune balance to repair the intestinal mucosal barrier.

[0029] Poria: sweet, mild and neutral, it enters the heart, spleen and kidney meridians, promotes diuresis and eliminates dampness, strengthens the spleen and calms the heart, and is mainly used to treat edema and oliguria. The triterpenoid components such as pachymic acid in the extract can enhance the phagocytic function of macrophages.

[0030] Preferably, the preparation method of the adsorbent comprises the following steps: taking volatile drug residues and extract drug residues, grinding to obtain drug residue powder; dispersing the drug residue powder into a starch solution, stirring and mixing evenly, to obtain a drug residue starch solution; adding calcium carbonate powder into the drug residue starch solution, stirring and mixing evenly, to obtain calcium carbonate with a surface coated with drug residue starch, roasting, and then pickling with hydrochloric acid to obtain the adsorbent;

[0031] Preferably, the calcium carbonate powder has a particle size of 0.5 to 2 mm.

[0032] First, the drug residue powder is mixed with a starch solution modified by phosphoric acid, and the phosphoric acid promotes the cross-linking of starch molecules to form a three-dimensional network skeleton; then, calcium carbonate powder is introduced as a template, and in the step-by-step temperature pyrolysis process, organic matter is carbonized to form a porous carbon matrix in the low-temperature stage, and calcium carbonate reacts with the carbon layer in a solid phase to generate active calcium oxide and release gas to form pores in the medium-temperature stage. In the high-temperature stage, calcium carbonate is completely decomposed to generate carbon dioxide, which reacts with the carbon matrix to gasify and expand the pores; finally, hydrochloric acid selectively etches calcium oxide to generate soluble salts, exposing hierarchical pores, and phosphoric acid activates the retained oxygen-containing functional groups to give the surface chemical adsorption activity, forming a composite adsorption material with both physical adsorption channels and chemical binding sites.

[0033] Preferably, the medicinal residue is ground to 500-1000 mesh; the starch solution is prepared by dissolving starch in a 10wt% phosphoric acid aqueous solution to obtain a starch solution with a concentration of 30-40wt%; the dosage ratio of the medicinal residue powder to the starch solution is 10g:15-30mL; the dosage ratio of the calcium carbonate powder to the medicinal residue starch solution is 10g:10-15mL; and the roasting conditions are pyrolysis at 400-600°C for 30-60min, pyrolysis at 600-800°C for 30-60min, and pyrolysis at 800-900°C for 30-60min.

[0034] Preferably, the penetrant is one or more of menthol, glycerin, water sap, lauryl azone, azone, propylene glycol, and ethanol.

[0035] Preferably, the excipient is one or more of vaseline, dimethicone, lanolin, and beeswax.

[0036] The present invention also claims protection for a method for preparing the Chinese medicine plaster for assisting the treatment of Clostridium difficile infection and restoring the balance of intestinal flora, comprising the following steps: mixing the active ingredient with an adsorbent, and then dispersing it in deionized water, adding a penetrant and an excipient, stirring and mixing evenly to obtain an ointment, applying the ointment on a substrate layer, and covering it with a protective layer to obtain the Chinese medicine plaster for assisting the treatment of Clostridium difficile infection and restoring the balance of intestinal flora.

[0037] The present invention also claims to protect the use of a traditional Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora, characterized in that it includes the following steps: cleaning the navel and the skin around it, keeping them dry, aligning the core part of the ointment with the center of the navel, applying it for 4 to 8 hours a day, 1 to 2 times a day, and continuously using it for 5 to 10 days.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] 1. The present invention provides a Chinese medicine patch for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora. The Chinese medicine patch has the advantages of administering the drug through the navel. The navel (Shenque point) is the intersection of meridians. After the drug penetrates through the skin, it can directly stimulate the acupuncture points to stimulate the meridian qi and regulate the circulation of gastrointestinal qi and blood through the Ren meridian. At the same time, its anatomical characteristics of thin epidermis and rich blood vessels significantly improve the bioavailability of the drug; the effective ingredients loaded by the adsorbent are sustained-released through the hollow porous structure, avoiding the problem of rapid dissipation of volatile oils in traditional patches; the penetrant reduces the skin barrier resistance by changing the lipid arrangement of the stratum corneum and promotes the penetration of small molecule drugs; the excipient provides suitable adhesion and ductility to ensure that the patch is in close contact with the skin and maintains continuous release of the drug.

[0040] 2. The present invention provides an ointment, wherein the active ingredients of Atractylodes macrocephala volatile oil help increase beneficial intestinal bacteria, and Poria cocos polysaccharide enhances the protective function of the flora and jointly inhibits the growth of harmful bacteria; the active ingredients of Magnolia officinalis and Artemisia argyi destroy the structure of Clostridium difficile and reduce toxin production, while the components of Coptis chinensis prevent bacterial division and reproduction; the extracts of White Peony Root and Atractylodes macrocephala promote tight connection of intestinal cells and repair mucosal barriers; the components of Dandelion and Magnolia officinalis reduce intestinal damage by inhibiting the inflammatory response pathway. Each component forms a linkage mechanism of "antibacterial-detoxification-repair-anti-inflammatory", which gradually takes effect from removing pathogens to restoring intestinal health.

[0041] 3. The present invention provides an adsorbent, calcium carbonate is pyrolyzed to produce CO 2 The formed microporous-mesoporous system greatly increases the drug loading capacity and can simultaneously encapsulate volatile oils and extracts; the active sites remaining in the Chinese medicine residues bind to the drug components through non-covalent bonds, and slow down the escape rate of volatile oils through the pore confinement effect, ensuring the stable release of the active ingredients throughout the entire use cycle. In addition, the resource utilization of drug residues combines environmental protection with optimized drug loading performance, which not only solves the problem of rapid volatilization of volatile oils in traditional plasters, but also prolongs the drug residence time by applying the natural closed environment of the umbilical depression, strengthening the synergistic effect of intestinal mucosal immune repair and flora regulation, and improving the treatment effect of Clostridium difficile infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show schematic diagrams of certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0043] Figure 1 The present invention is a Chinese medicine plaster for treating Clostridium difficile infection and the survival rate of mice for reconstructing the balance of intestinal flora. DETAILED DESCRIPTION

[0044] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with embodiments. Of course, the specific embodiments described here are only used to explain the present invention and are not used to limit the present invention.

[0045] Unless otherwise specified, the chemical reagents and materials in the present invention are purchased from commercial sources or synthesized from raw materials purchased from commercial sources.

[0046] A method for preparing a traditional Chinese medicine plaster for assisting in treating Clostridium difficile infection and restoring intestinal flora balance comprises the following steps:

[0047] (1) taking Atractylodes macrocephala, Magnolia officinalis, and Artemisia argyi, crushing them into 40-80 meshes, adding them into deionized water 10-20 times the weight of the traditional Chinese medicine and soaking them for 30-60 minutes, refluxing and extracting them at 100° C., collecting volatile oils, respectively obtaining the Atractylodes macrocephala volatile oil, Magnolia officinalis volatile oil, and Artemisia argyi volatile oil, and obtaining volatile drug residues for later use;

[0048] (2) taking coptis root, dandelion, white peony root and poria, adding them to 60-80wt% ethanol which is 8-15 times the weight of the traditional Chinese medicine, heating and refluxing extraction for 2 times, each time for 1-2h, combining the extracts, filtering, decompressing the filtrate to recover ethanol, and concentrating it into a thick paste, respectively obtaining the coptis root extract, dandelion extract, white peony root extract and poria extract, and obtaining the extract residue for standby use;

[0049] (3) Grinding the volatile drug residue and the extract drug residue to 500-1000 mesh to obtain drug residue powder; dispersing 10 g of the drug residue powder into 15-30 mL of a 30-40 wt% starch solution (obtained by dissolving starch in a 10 wt% phosphoric acid aqueous solution), stirring and mixing to obtain a drug residue starch solution; adding 10 g of calcium carbonate powder into 10-15 mL of the drug residue starch solution, stirring and mixing to obtain calcium carbonate coated with drug residue starch, pyrolyzing at 400-600° C. for 30-60 min, 600-800° C. for 30-60 min, and 800-900° C. for 30-60 min, and then washing with hydrochloric acid to obtain the adsorbent;

[0050] (4) Take 4-5 parts of Atractylodes macrocephala volatile oil, 3-6 parts of Magnolia officinalis volatile oil, 1-3 parts of Artemisia argyi volatile oil, 16-24 parts of Coptis chinensis extract, 8-12 parts of Taraxacum officinale extract, 12-18 parts of White Peony Root extract, and 12-20 parts of Poria cocos extract to obtain effective ingredients; mix 1-6 wt% of the effective ingredients with 40-52 wt% of the adsorbent, and then disperse them in 38-58 wt% of deionized water, add 0.5-2 wt% of the penetrant and 0.5-2 wt% of the excipient, stir and mix evenly to obtain an ointment, apply the ointment on the substrate layer, and cover it with a protective layer to obtain the Chinese medicine plaster for auxiliary treatment of Clostridium difficile infection and restoration of intestinal flora balance.

[0051] The penetrant is one or more of menthol, glycerin, water saturated fatty acid, lauryl azone, azone, propylene glycol, and ethanol; the excipient is one or more of vaseline, dimethicone, lanolin, and beeswax.

[0052] The present invention will be further described below through specific embodiments.

[0053] Example 1

[0054] A method for preparing a traditional Chinese medicine plaster for assisting in treating Clostridium difficile infection and restoring intestinal flora balance comprises the following steps:

[0055] (1) taking Atractylodes macrocephala, Magnolia officinalis, and Artemisia argyi, crushing them into 50 mesh, adding them into deionized water 15 times the weight of the traditional Chinese medicine and soaking them for 45 minutes, refluxing and extracting them at 100° C., collecting volatile oils, respectively obtaining the Atractylodes macrocephala volatile oil, Magnolia officinalis volatile oil, and Artemisia argyi volatile oil, and obtaining volatile drug residues for later use;

[0056] (2) taking coptis root, dandelion, white peony root and poria, adding them to 70wt% ethanol which is 12 times the weight of the traditional Chinese medicine, heating and refluxing extraction for 2 times, each time for 1.5h, combining the extracts, filtering, decompressing the filtrate to recover ethanol, and concentrating it into a thick paste, respectively obtaining the coptis root extract, dandelion extract, white peony root extract and poria extract, and obtaining the extract residue for standby use;

[0057] (3) Grinding the volatile drug residue and the extract drug residue to 800 mesh to obtain drug residue powder; dispersing 10 g of the drug residue powder into 25 mL of a 35 wt% starch solution (obtained by dissolving starch in a 10 wt% phosphoric acid aqueous solution), stirring and mixing to obtain a drug residue starch solution; adding 10 g of calcium carbonate powder to 12 mL of the drug residue starch solution, stirring and mixing to obtain calcium carbonate coated with drug residue starch, pyrolyzing at 500° C. for 45 min, 700° C. for 45 min, and 850° C. for 45 min, and then washing with hydrochloric acid to obtain the adsorbent;

[0058] (4) Take 4.5 g of Atractylodes macrocephala volatile oil, 4.5 g of Magnolia officinalis volatile oil, 2 g of Artemisia argyi volatile oil, 20 g of Coptis chinensis extract, 10 g of Taraxacum officinale extract, 15 g of White Peony Root extract, and 16 g of Poria cocos extract to obtain effective ingredients; mix 3.5 g of the effective ingredients with 46 g of the adsorbent, and then disperse them in 48 g of deionized water, add 1 g of the penetrant laurocapron and 1.5 g of the excipient beeswax, stir and mix evenly to prepare an ointment, apply the ointment on a non-woven fabric substrate layer with a diameter of 2 to 5 cm, and cover it with a non-woven fabric protective layer to obtain the Chinese medicine plaster for auxiliary treatment of Clostridium difficile infection and restoration of intestinal flora balance.

[0059] Comparative Example 1

[0060] A method for preparing a traditional Chinese medicine plaster for assisting in treating Clostridium difficile infection and restoring intestinal flora balance comprises the following steps:

[0061] (1) taking Atractylodes macrocephala, Magnolia officinalis, and Artemisia argyi, crushing them into 50 mesh, adding them into deionized water 15 times the weight of the traditional Chinese medicine and soaking them for 45 minutes, refluxing and extracting them at 100° C., collecting volatile oils, respectively obtaining the Atractylodes macrocephala volatile oil, Magnolia officinalis volatile oil, and Artemisia argyi volatile oil, and obtaining volatile drug residues for later use;

[0062] (2) taking coptis root, dandelion, white peony root and poria, adding them to 70wt% ethanol which is 12 times the weight of the traditional Chinese medicine, heating and refluxing extraction for 2 times, each time for 1.5h, combining the extracts, filtering, decompressing the filtrate to recover ethanol, and concentrating it into a thick paste, respectively obtaining the coptis root extract, dandelion extract, white peony root extract and poria extract, and obtaining the extract residue for standby use;

[0063] (3) grinding the volatile drug residue and the extract drug residue to 800 mesh to obtain drug residue powder; pyrolyzing 10 g of the drug residue powder at 500° C. for 45 min, 700° C. for 45 min, and 850° C. for 45 min, and then washing with hydrochloric acid to obtain the adsorbent;

[0064] (4) Take 4.5 g of Atractylodes macrocephala volatile oil, 4.5 g of Magnolia officinalis volatile oil, 2 g of Artemisia argyi volatile oil, 20 g of Coptis chinensis extract, 10 g of Taraxacum officinale extract, 15 g of White Peony Root extract, and 16 g of Poria cocos extract to obtain effective ingredients; mix 3.5 g of the effective ingredients with 46 g of the adsorbent, and then disperse them in 48 g of deionized water, add 1 g of the penetrant laurocapron and 1.5 g of the excipient beeswax, stir and mix evenly to prepare an ointment, apply the ointment on a non-woven fabric substrate layer with a diameter of 2 to 5 cm, and cover it with a non-woven fabric protective layer to obtain the Chinese medicine plaster for auxiliary treatment of Clostridium difficile infection and restoration of intestinal flora balance.

[0065] Experiment on the treatment of diarrhea in mice with Chinese medicine plaster.

[0066] 1. Experimental Materials

[0067] 1. Experimental Animals

[0068] SPF KM mice (6 weeks old, weight 20±2g), 10 mice per group, half male and half female, 4 groups in total (normal group, infection group, Example 1 group, Comparative Example 1 group)

[0069] 2. Strains and reagents

[0070] Escherichia coli ATCC25922 (bacterial concentration 1×109 CFU / mL);

[0071] Antibiotic mixture (gentamicin sulfate 65 g / L + cefradine 65 g / L);

[0072] Embodiment 1 Chinese medicine plaster;

[0073] Comparative Example 1 Chinese medicine plaster.

[0074] 2. Experimental Methods

[0075] 1. Establishment of animal model

[0076] Antibiotic pretreatment: The mice in the infection group, Example 1 group, and Comparative Example 1 group were intragastrically administered with 25 mL / (kg·d) of antibiotic mixture every day for 5 consecutive days to destroy the intestinal flora.

[0077] Pathogen infection: After pretreatment, 0.2 mL (1×109 CFU / mL) of Escherichia coli ATCC25922 solution was gavaged and the rats were observed for 24 hours until symptoms of lethargy, loose stools, and perianal contamination appeared.

[0078] 2. Grouping and Dosing

[0079] Normal group: no infection, no intervention.

[0080] Infection group: no treatment after infection.

[0081] Group 1 of Example 1: 24 hours after infection, the patch of Example 1 was applied to the navel of the mice (changed once a day for 7 days).

[0082] Comparative Example 1 Group: Same as Example 1 Group, except that the Comparative Example 1 patch was applied.

[0083] 3. Observation indicators

[0084] Feces score: The size of the stain area formed by feces on the filter paper is graded into 4 levels: Level 1 is a contamination diameter of less than 1.0 cm, indicated by "+"; Level 2 is a contamination diameter of 1.0-1.9 cm, indicated by "++"; Level 3 is a contamination diameter of 2.0-3.0 cm, indicated by "+++"; Level 4 is a contamination diameter of more than 3.0 cm, indicated by "++++". The specific results are shown in Table 1.

[0085] Table 1 Fecal scoring results

[0086] Normal group Infection group Example 1 Comparative Example 1 Fecal score after modeling + ++++ ++++ ++++ Fecal score after treatment + ++++ + ++

[0087] Diarrhea rate: According to "Pharmacological Experimental Methodology" (4th edition), the diarrhea rate on the 0th, 3rd and 7th day of the experiment was measured respectively. Diarrhea rate = number of diarrheal animals / total number in the group × 100%. The specific results are shown in Table 2.

[0088] Table 2 Diarrhea rate test results

[0089]

[0090] Inflammatory factors: 24 hours after the last administration, serum and small intestinal tissues were collected from mice in each group, and ELISA was used to detect the levels of TNF-α and IL-6. The specific data are shown in Table 3.

[0091] Table 3 Inflammatory factor detection results

[0092]

[0093] Note: * indicates p < 0.05 compared with the normal group; # indicates p < 0.05 compared with the infection group.

[0094] Therapeutic experiment of traditional Chinese medicine plaster on Clostridium difficile-infected mice.

[0095] I. Experimental materials

[0096] 1. Experimental animals

[0097] SPF-grade BALB / c mice (6 weeks old, male, body weight 20 ± 2 g), 10 mice in each group, a total of 5 groups (normal group, infection group, Example 1 group, Comparative Example 1 group, vancomycin group).

[0098] 2. Strains and reagents

[0099] Clostridium difficile ATCC43255 (anaerobic culture, bacterial liquid concentration 1 × 109 CFU / mL);

[0100] Antibiotic mixture (kanamycin 0.4 mg / mL, gentamicin 0.035 mg / mL, colistin 850 U / mL, metronidazole 0.215 mg / mL, vancomycin 0.045 mg / mL);

[0101] Clindamycin injection (10 mg / kg);

[0102] Vancomycin suspension (50 mg / kg, oral);

[0103] Traditional Chinese medicine plaster of Example 1;

[0104] Traditional Chinese medicine plaster of Comparative Example 1.

[0105] II. Experimental methods

[0106] 1. Establishment of animal model

[0107] Antibiotic pretreatment: Mice were allowed to drink water containing the antibiotic mixture freely for 5 days, and then changed to sterilized water for 3 days.

[0108] Infection induction: Intraperitoneal injection of clindamycin (10 mg / kg), and 24 hours later, 0.2 mL of Clostridium difficile ATCC43255 bacterial liquid was administered by gavage.

[0109] 2. Grouping and administration

[0110] Normal group: No infection, no intervention.

[0111] Infection group: Not treated after infection.

[0112] Example 1 group: 16 hours after infection, the plaster of Example 1 was applied at the navel (changed once a day for 5 days).

[0113] Control Example 1 group: The same as Example 1 group, apply the plaster of Control Example 1.

[0114] Vancomycin group: Intragastric administration of vancomycin suspension (50 mg / kg, once a day, for 5 consecutive days) 16 hours after infection.

[0115] 3. Observation indicators

[0116] Survival rate: Record the death situation within 7 days. Specific data are shown in Figure 1 .

[0117] The above is only the preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, makes equivalent substitutions or changes, and should be covered by the protection scope of the present invention.

Claims

1. A Chinese medicine plaster for assisting the treatment of Clostridium difficile infection and restoring the balance of intestinal flora, comprising a substrate layer, an ointment, and a protective layer, characterized in that: The ointment is prepared from 1-6wt% of active ingredients, 0.5-2wt% of penetrant, 0.5-2wt% of excipient, 40-52wt% of adsorbent, and 38-58wt% of deionized water; the active ingredients are prepared from the following components in parts by weight: 4-5 parts of Atractylodes macrocephala volatile oil, 3-6 parts of Magnolia officinalis volatile oil, 1-3 parts of Artemisia argyi volatile oil, 16-24 parts of Coptis chinensis extract, 8-12 parts of Taraxacum officinale extract, 12-18 parts of White Peony Root extract, and 12-20 parts of Poria cocos extract.

2. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 1, characterized in that: The preparation method of Chinese medicine volatile oil is as follows: take Atractylodes macrocephala, Magnolia officinalis, and Artemisia argyi, crush them, add them into deionized water for soaking, reflux extraction, collect volatile oil, respectively obtain the Atractylodes macrocephala volatile oil, Magnolia officinalis volatile oil, Artemisia argyi volatile oil, and obtain volatile drug residues for standby use.

3. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 2, characterized in that: The Chinese medicine is crushed into 40-80 meshes, the weight of deionized water is 10-20 times of the Chinese medicine, soaked for 30-60 minutes, and the reflux extraction temperature is 100°C.

4. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 1, characterized in that: The preparation method of the Chinese medicine extract is as follows: take coptis root, dandelion, white peony root and poria, add them to ethanol, heat and reflux, combine the extracts, filter, reduce the pressure of the filtrate to recover the ethanol, and concentrate it into a thick paste to obtain the coptis root extract, dandelion extract, white peony root extract and poria extract respectively, and obtain the extract residue for later use.

5. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 4, characterized in that: The concentration of ethanol is 60-80 wt %, and the weight of ethanol is 8-15 times of the traditional Chinese medicine; the extraction is carried out by heating and reflux for 2 times, each time for 1-2 hours.

6. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 1, characterized in that: The preparation method of the adsorbent comprises the following steps: taking volatile drug residues and extract drug residues, grinding to obtain drug residue powder; dispersing the drug residue powder into a starch solution, stirring and mixing evenly to obtain a drug residue starch solution; adding calcium carbonate powder into the drug residue starch solution, stirring and mixing evenly to obtain calcium carbonate with drug residue starch coated on the surface, roasting, and then pickling with hydrochloric acid to obtain the adsorbent.

7. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 6, characterized in that: The medicinal residue is ground to 500-1000 meshes; the starch solution is prepared by dissolving starch in a 10wt% phosphoric acid aqueous solution to obtain a starch solution with a concentration of 30-40wt%; the dosage ratio of the medicinal residue powder to the starch solution is 10g:15-30mL; the dosage ratio of the calcium carbonate powder to the medicinal residue starch solution is 10g:10-15mL; the roasting conditions are pyrolysis at 400-600°C for 30-60min, pyrolysis at 600-800°C for 30-60min, and pyrolysis at 800-900°C for 30-60min.

8. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 1, characterized in that: The penetrant is one or more of menthol, glycerin, water sap, lauryl azone, azone, propylene glycol, and ethanol.

9. The Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora according to claim 1, characterized in that: The excipient is one or more of vaseline, dimethicone, lanolin and beeswax.

10. A method for preparing the Chinese medicine plaster for assisting in treating Clostridium difficile infection and restoring the balance of intestinal flora as claimed in any one of claims 1 to 9, characterized in that: The method comprises the following steps: mixing the active ingredient with the adsorbent, dispersing the mixture into deionized water, adding a penetrant and an excipient, stirring and mixing the mixture evenly to obtain an ointment, applying the ointment onto a substrate layer, and covering the ointment with a protective layer to obtain the Chinese medicine plaster for assisting in the treatment of Clostridium difficile infection and restoring the balance of intestinal flora.

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